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<i>Euonymus</i> species from the Celastraceae family are considered as a source of unusual genes modifying the oil content and fatty acid composition of vegetable oils. Due to the possession of genes encoding enzyme diacylglycerol acetyltransferase (DAcT), <i>Euonymus</i> plants can synthesize and accumulate acetylated triacyglycerols. The gene from <i>Euonymus europaeus</i> (<i>EeDAcT</i>) encoding the DAcT was identified, isolated, characterized, and modified for cloning and genetic transformation of plants. This gene has a unique nucleotide sequence and amino acid composition, different from orthologous genes from other <i>Euonymus</i> species. Nucleotide sequence of original <i>EeDAcT</i> gene was modified, cloned into transformation vector, and introduced into tobacco plants. Overexpression of <i>EeDAcT</i> gene was confirmed, and transgenic host plants produced and accumulated acetylated triacylglycerols (TAGs) in immature seeds. Individual transgenic plants showed difference in amounts of synthesized acetylTAGs and also in fatty acid composition of acetylTAGs.